• Vaccine Ontology ID: VO_0011464
• Type: Subunit vaccine
• Status: Research
• Antigen: Canine parvovirus VP2
• VP2 gene engineering:
  • Type: Recombinant protein preparation
  • Description: CPMV–PARVO1 contains a 17-amino acid peptide (DGAVQPDGGQPAVRNER) corresponding to residues 3 to 19 from VP2 of canine parvovirus, inserted between residues 22 and 23 of the S protein of CPMV. Each virion contains 60 copies of this peptide and 1 μg chimera contains approximately 20 ng of peptide. The construction, propagation and purification of CPMV–PARVO1 virions (and wild type CPMV) was carried out. In addition, the virus particles were inactivated by irradiation with UV254 light during the purification, according to the following procedure: after resuspension of the PEG/NaCl pellet and clarification, the concentration of particles was measured by spectrophotometry, and the virus-containing supernatant diluted to 1 mg/ml (Langeveld et al., 2001).
  • Detailed Gene Information: Click Here.
• Immunization Route: Subcutaneous injection

• Vaccine Ontology ID: VO_0011462
• Type: DNA vaccine
• Status: Research
• Antigen: Canine parvovirus VP1
• VP1 gene engineering:
  • Type: DNA vaccine construction
  • Description: CPV viral DNA was used as template to amplify the entire VP1 fragment, including VP2, by polymerase chain reaction (PCR). Two deoxyoligonucleotides, coding and noncoding strand primers were synthesized. The product was then electrophoresed in a 0.8% agarose/TAE gel. The band containing the PCR product was excised from the agarose gel by QIAGEN QIAquickTM spin columns (QIAGEN GmbH, Hilden, Germany) and digested simultaneously with BamHI and Not1 (New England Biolabs, Beverly, MA, USA). The digested VP1 fragment was ligated into BamHIINotI digested pGT36 (kindly provided by Dr David Curiel, University of Alabama, Birmingham, AL, USA), a derivative of pCDNA3, which has a CMV promoter and kanamycin resistance gene. Kanamycin resistance clones were screened for appropriate restriction digestion patterns and one clone, designated as pGT36VP1, was then sequenced using an Applied Biosystems Model 373A Automated Sequencer (Perkin-Elmer/Applied Biosysterns Division, Folster City, CA, USA) (Jiang et al., 1998).
  • Detailed Gene Information: Click Here.
• Vector: plasmid vector pGT36
• Immunization Route: Intramuscular injection (i.m.)

Dog Response

• Vaccination Protocol: Dogs were kept in four groups of five or six animals, A–D, each group in a separate room. Groups A, B and C (six dogs in each) were immunized subcutaneously above the shoulder blades on days 0 and 28 with either CPMV–PARVO1, 3L17–KLH or wild type CPMV, respectively, in Quil-A®/Alhydrogel®. A fourth group (D) of five dogs was left unvaccinated. Inoculation sites were checked daily for adverse reactions (Langeveld et al., 2001).
• Challenge Protocol: On day 42, all 23 dogs were challenged with CPV by the oronasal route [19], by distributing CPV onto their eyes, noses and mouths (1.5 ml/dog). Blood was collected prior to immunization, following immunization (days 28 and 38) and following challenge (day 48 and 57), and sera collected and stored at −20°C. Rectal swabs were performed with cotton wool swab sticks and stored at −20°C prior to analysis for the presence of CPV (Langeveld et al., 2001).
• Efficacy: Dogs immunized with the inactivated CPMV-PARVO1 in adjuvant displayed no clinical signs of disease and shedding of CPV in faeces was limited following CPV challenge (Langeveld et al., 2001).

Dog Response

• Vaccination Protocol: Four mix breed dogs, 9 months of age, from a single litter were injected with doses of 200, 400, 600 or 800 /lg pGT36VPl in a total volume of 1 ml phosphate buffered saline (PBS, 0.067 M, pH 7.2). One negative control sibling was injected with 1 ml PBS. All injections were administered i.m. at the right quadriceps muscle. Blood samples were collected weekly. All dogs were reinjected on week 16 with the same dose of DNA used initially at the same site and the negative control was injected with PBS (Jiang et al., 1998).
• Challenge Protocol: Challenge infections with virulent CPV were performed in a HEIPA filtered, environmentally controlled kennel facility at the College of Veterinary Medicine, Auburn University. The challenge strains of CPV (AU-H3-WS-93) consist of equal amounts of three virulent CPV slrains, AU-H3 (CPV-2), WS (CPV-2a) and 93 (CPV-2b) at a dose of 10^5 TCID(50) (tissue culture infectious dose) per ml. All challenge strains were purified from fecal extracts. The challenge dose was given intranasally (0.5 ml in each nostril) and orally (1.0 ml). The dogs were monitored three times daily for clinical signs including attitude, appetite, stool consistency, temperature and vomiting (Jiang et al., 1998).
• Efficacy: All pGT36VP1 vaccinated dogs were protected against infection after virulent CPV challenge regardless of dose and the unvaccinated control dog was fully susceptible (Jiang et al., 1998).
• Host IgG Fc fragment response
  • Description: NAV vaccinated dogs showed an increase of serum IgG titer starting 1 week post-injection which peaked at week 2 and remained detectable for at least 14 weeks. The non-immunized control dog did not show an antibody response at any point pre-challenge (Jiang et al., 1998).
  • Detailed Gene Information: Click Here.